Infection and transmission of SARS-CoV-2 depends on heparan sulfate proteoglycans

  1. Marta Bermejo-Jambrina
  2. Julia Eder
  3. Tanja M. Kaptein
  4. John L. van Hamme
  5. Leanne C. Helgers
  6. Killian E. Vlaming
  7. Philip J.M. Brouwer
  8. Alexander P.J. Vlaar
  9. Frank E.H.P. van Baarle
  10. Marcel Spaargaren
  11. Godelieve J. de Bree
  12. Bernadien M. Nijmeijer
  13. Neeltje A. Kootstra
  14. Marit J. van Gils
  15. Rogier W. Sanders
  16. Teunis B. H. Geijtenbeek

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Abstract

The current pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and outbreaks of new variants highlight the need for preventive treatments. Here we identified heparan sulfate proteoglycans as attachment receptors for SARS-CoV-2. Notably, neutralizing antibodies against SARS-CoV-2 isolated from COVID-19 patients interfered with SARS-CoV-2 binding to heparan sulfate proteoglycans, which might be an additional mechanism of antibodies to neutralize infection. SARS-CoV-2 binding to and infection of epithelial cells was blocked by low molecular weight heparins (LMWH). Although dendritic cells (DCs) and mucosal Langerhans cells (LCs) were not infected by SARS-CoV-2, both DC subsets efficiently captured SARS-CoV-2 via heparan sulfate proteoglycans, and transmitted the virus to ACE2-positive cells. Moreover, human primary nasal cells were infected by SARS-CoV-2 and infection was blocked by pre-treatment with LMWH. These data strongly suggest that heparan sulfate proteoglycans are important attachment receptors facilitating infection and transmission, and support the use of LMWH as prophylaxis against SARS-CoV-2 infection.

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  1. ScreenIT

    SciScore for 10.1101/2020.08.18.255810: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board StatementIRB: Alveolar macrophages were prepared from broncheo-alveolar lavage (BAL) fluid that was obtained as spare material from the ongoing DIVA study (Netherlands Trial Register: NL6318; AMC Medical Ethical Committee approval number: 2014_294).
    Consent: All subjects in the DIVA study have signed an informed consent form.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableThe DIVA study includes healthy male volunteers aged 18-35.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Reagents and antibodies: The following antibodies were used (all anti-human): ACE-2 (R&D),
    anti-human): ACE-2
    suggested: None
    Negative control included isotype-matched HIV-1 antibody VRC01 (59)
    HIV-1
    suggested: (bNAber Cat# bNAberID_1, RRID:AB_2491019)
    Purity of LCs was routinely verified by flow cytometry using antibodies directed against CD207 (langerin) and CD1a.
    CD207
    suggested: None
    CD1a
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Cell lines: The human B cell line Namalwa (ATCC, CRL-1432) and Namalwa cells stably expressing human Syndecan 1 and Syndecan 4 (57) were a gift from Dr. Guido David and Dr. Philippe A Gallay.
    Namalwa
    suggested: None
    The human epithelial Caco2 cells (ATCC, HTB-37™) were maintained in Dulbecco modified Eagle medium (Gibco Life Technologies, Gaithersburg, Md.) containing 10% fetal calf serum (FCS), L-glutamine and penicillin/streptomycin (10 μg/ml) and supplemented with MEM Non-Essential Amino Acids Solution (NEAA) (Gibco Life Technologies, Gaithersburg, Md.).
    Caco2
    suggested: ATCC Cat# HTB-37, RRID:CVCL_0025)
    Biosynthesis inhibition and enzymatic treatment: HuH7.5 cells were treated in D-PBS/0.25% BSA with 46 miliunits heparinase III (Amsbio) for 1 hour at 37°C, washed and used in subsequent experiments.
    HuH7.5
    suggested: RRID:CVCL_7927)
    Software and Algorithms
    SentencesResources
    Data was analyzed using FlowJo vX.
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    Statistical analyses were performed using GraphPad Prism 8 software and significance was set at *P< 0.05, **P<0.01***P<0.001****P<0.0001.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. Version published to 10.1101/2020.08.18.255810 on bioRxiv